sequence and linker dependent chiral dimerization of dna–porphyrin conjugates
TRANSCRIPT
Supporting information for the manuscript
Sequence and linker dependent chiral dimerization of DNA-
porphyrin conjugates
Gevorg Sargsyan, Brianna L. MacLeod, Urice Tohgha, and Milan Balaz*
University of Wyoming, Department of Chemistry, 1000 E. University Ave, Laramie, WY
82071, USA. phone: +1 307 761-1657, email: [email protected]
List of contents
MALDI-TOF MS spectra of DNA-porphyrin conjugates ……………………………… p. S1-S4
HPLC analysis of the DNA-porphyrin conjugates ……………………………… p. S5-S7
UV-vis studies of the DNA-porphyrin conjugates ……………………………… p. S8-S15
Fluorescence studies of the DNA-porphyrin conjugates ……………………………… p. S16-S22
RLS studies of the DNA-porphyrin conjugates ……………………………… p. S23-S24
CD studies of the DNA-porphyrin conjugates ……………………………… p. S25-S29
S1
MADI-TOF MS of DNA-porphyrin conjugates
Figure S1. MALDI-TOF MS of 2H-aA m/z MALDI-TOF 3085.7768 (C122H123N48O38P7, [M-H]+
requires
3085.7409).
Figure S2. MALDI-TOF MS of Zn-aA m/z MALDI-TOF 3147.6621 (C122H121N48O38P7Zn, [M-H]+
requires 3147.6544).
S2
Figure S3. MALDI-TOF MS of 2H-pA m/z MALDI-TOF 3152.7562 (C122H125N47O41P8, [M-H]+
requires
3152.7120).
Figure S4. MALDI-TOF MS of Zn-pA m/z MALDI-TOF 3214.6550 (C122H123N47O41P8Zn, [M-H]+
requires 3214.6255).
S3
Figure S5. MALDI-TOF MS of 2H-aT m/z MALDI-TOF 3013.6120 (C122H131N24O54P7, [M-H]+
requires
3013.6484).
Figure S6. MALDI-TOF MS of Zn-aT m/z MALDI-TOF 3075.5914 (C122H129N24O54P7Zn, [M-H]+
requires 3075.5619).
S4
Figure S7. MALDI-TOF MS of 2H-pT m/z MALDI-TOF 3080.6127 (C122H133N23O57P8, [M-H]+
requires
3080.6194).
Figure S8. MALDI-TOF MS of Zn-pT m/z MALDI-TOF 3142.5241 (C122H132N23O57P8Zn, [M-H]+
requires 3142.5329).
S5
HPLC analysis of the DNA-porphyrin conjugates
HPLC analysis of synthesized and purified porphyrin-ODN conjugates was carried out on a Hitachi
LaChrom ELITE HPLC system equipped with L-2130 quaternary pump, L-2455 diode array detector, L-
2200 autosampler, and L-2350 column oven using X-Terra analytical column MS C18 2.5µm 4.6 × 50
mm at 55 °C, 1 mL / min flow.
Minutes
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18
mA
U
0
100
200
300
mA
U
0
100
200
300
DAD-CH3 420 - 700 nm
An-2H-aA8-55C-
Figure S9. HPLC chromatogram of the DNA-porphyrin conjugate 2H-aA8 monitored at 420 nm. Inlet:
UV-vis absorption profile of the peak recorded by PDA detector.
Minutes
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18
mA
U
0
100
200
300
400
mA
U
0
100
200
300
400
DAD-CH3 420 - 700 nm
An-2H-pT8-55C-
Figure S10. HPLC chromatogram of the DNA-porphyrin conjugate 2H-pT8 monitored at 420 nm. Inlet:
UV-vis absorption profile of the peak recorded by PDA detector.
nm
300 400 500 600 700
mA
U
0
100
200
300
400
500
mA
U
0
100
200
300
400
50010.27 Min
An-2H-aA8-55C-
nm
300 400 500 600 700
mA
U
0
200
400
600
mA
U
0
200
400
600
10.13 Min
An-2H-pT8-55C-
S6
Minutes
0 2 4 6 8 10 12 14 16 18 20 22
mA
U
0
50
100
150
200
mA
U
0
50
100
150
200
DAD-CH3 420 - 700 nm
An-2H-aT8-55C-
Figure S11. HPLC chromatogram of the DNA-porphyrin conjugate 2H-aT8 monitored at 420 nm. Inlet:
UV-vis absorption profile of the peak recorded by PDA detector.
Minutes
0 2 4 6 8 10 12 14 16 18 20 22
mA
U
0
50
100
150
mA
U
0
50
100
150
DAD-CH3 420 - 700 nm
An-2H-aA8-55C-
Figure S12. HPLC chromatogram of the DNA-porphyrin conjugate 2H-aA8 monitored at 420 nm. Inlet:
UV-vis absorption profile of the peak recorded by PDA detector.
Minutes
0 2 4 6 8 10 12 14 16 18 20 22
mA
U
0
20
40
60
80m
AU
0
20
40
60
80
DAD-CH3 420 - 700 nm
An-Zn-aA8-55C-
Figure S13. HPLC chromatogram of the DNA-porphyrin conjugate Zn-aA8 monitored at 420 nm. Inlet:
UV-vis absorption profile of the peak recorded by PDA detector.
nm
300 400 500 600 700
mA
U
0
50
100
150
200
250
mA
U
0
50
100
150
200
25010.32 Min
An-2H-aT8-55C-
nm
300 400 500 600 700
mA
U
0
20
40
60
80
100
mA
U
0
20
40
60
80
100
10.32 Min
An-Zn-aA8-55C-
nm
300 400 500 600 700
mA
U
0
50
100
150
200
mA
U
0
50
100
150
200
10.28 Min
An-2H-aA8-55C-
S7
Minutes
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18
mA
U
0
50
100
150
200
mA
U
0
50
100
150
200DAD-CH3 420 - 700 nm
An-Zn-pT8-55C-
Figure S14. HPLC chromatogram of the DNA-porphyrin conjugate Zn-pT8 monitored at 420 nm. Inlet:
UV-vis absorption profile of the peak recorded by PDA detector.
Minutes
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18
mA
U
0
100
200
mA
U
0
100
200
DAD-CH3 420 - 700 nm
An-Zn-aT8-55C-
Figure S15. HPLC chromatogram of the DNA-porphyrin conjugate Zn-aT8 monitored at 420 nm. Inlet:
UV-vis absorption profile of the peak recorded by PDA detector.
Minutes
0 2 4 6 8 10 12 14 16 18 20 22
mA
U
0
50
100
150
200m
AU
0
50
100
150
200DAD-CH3 420 - 700 nm
An-Zn-pA8-55C-
Figure S16. HPLC chromatogram of the DNA-porphyrin conjugate Zn-pA8 monitored at 420 nm. Inlet:
UV-vis absorption profile of the peak recorded by PDA detector.
nm
300 400 500 600 700
mA
U
0
50
100
150
200
mA
U
0
50
100
150
200
10.10 Min
An-Zn-pT8-55C-
nm
300 400 500 600 700m
AU
0
100
200
300
mA
U
0
100
200
30010.29 Min
An-Zn-aT8-55C-
nm
300 400 500 600 700
mA
U
0
50
100
150
200
250
mA
U
0
50
100
150
200
25010.08 Min
An-Zn-pA8-55C-
S8
UV-vis studies of the DNA-porphyrin conjugates
UV-vis temperature dependent experiments were carried out in 1 mM Na-cacodylate buffer, pH=7 with
different concentrations of NaCl. Temperature ramping rate was 1 °C/min.
Figure S18. Temperature dependent UV-vis absorption spectra of 2H-aA conjugate in the absence of
NaCl.
Figure S17. UV-vis absorption spectra of all eight DNA-porphyrin conjugates in 1 mM Na-cacodylate buffer,
S9
Figure19. Temperature dependent UV-vis absorption spectra of 2H-aA conjugate in the presence of 20
mM NaCl.
Figure S20. Temperature dependent UV-vis absorption spectra of 2H-aA conjugate in the presence of 40
mM NaCl.
S10
Figure S21. Temperature dependent UV-vis absorption spectra of 2H-aA conjugate in the presence of 60
mM NaCl.
Figure S22. Temperature dependent UV-vis absorption spectra of 2H-aA conjugate in the presence of 80
mM NaCl.
S11
Figure S23. Temperature dependent UV-vis absorption spectra of 2H-aA conjugate in the presence of
100 mM NaCl.
Figure S24. Temperature dependent UV-vis absorption spectra of Zn-aA conjugate in the absence of
NaCl.
S12
Figure S25. Temperature dependent UV-vis absorption spectra of Zn-aA conjugate in the presence of 20
mM NaCl.
Figure S26. Temperature dependent UV-vis absorption spectra of Zn-aA conjugate in the presence of 40
mM NaCl.
S13
Figure S27. Temperature dependent UV-vis absorption spectra of Zn-aA conjugate in the presence of 60
mM NaCl.
Figure S28. Temperature dependent UV-vis absorption spectra of Zn-aA conjugate in the presence of 80
mM NaCl.
S14
Figure 29. Temperature dependent UV-vis absorption spectra of Zn-aA conjugate in the presence of 100
mM NaCl.
Figure S30. Temperature dependent UV-vis absorption spectra of 2H-pA conjugate in the presence of 0
mM NaCl.
S15
Figure S31. Temperature dependent UV-vis absorption spectra of 2H-pA conjugate in the presence of
100 mM NaCl.
Figure S32. Temperature dependent UV-vis absorption spectra of 2H-pA conjugate in the presence of
500 mM NaCl.
.
S16
Fluorescence studies of the DNA-porphyrin conjugates
Temperature dependent fluorescence experiments were carried out in 1 mM Na-cacodylate buffer, pH=
7.0 with different concentrations of NaCl. Temperature ramping rate was 1 °C/min. The excitation and
emission slits were 2.5 nm.
Figure S33. Temperature dependent fluorescence spectra of 2H-aA conjugate in the absence of NaCl.
Figure S34. Temperature dependent fluorescence spectra of 2H-aA conjugate in the absence of NaCl.
S17
Figure S35. Temperature dependent fluorescence spectra of 2H-aA conjugate in the presence of 20 mM
NaCl.
Figure S36. Temperature dependent fluorescence spectra of 2H-aA conjugate in the presence of 20 mM
NaCl.
S18
Figure S37. Temperature dependent fluorescence spectra of 2H-aA conjugate in the presence of 40 mM
NaCl.
Figure S38. Temperature dependent fluorescence spectra of 2H-aA conjugate in the presence of 40 mM
NaCl.
S19
Figure S39. Temperature dependent fluorescence spectra of 2H-aA conjugate in the presence of 100 mM
NaCl.
Figure S40. Temperature dependent fluorescence spectra of 2H-aA conjugate in the presence of 100 mM
NaCl.
S20
Figure S41. Temperature dependent fluorescence spectra of Zn-aA conjugate in the absence of NaCl.
Figure S42. Temperature dependent fluorescence spectra of Zn-aA conjugate in the absence of NaCl.
S21
Figure S43. Temperature dependent fluorescence spectra of Zn-aA conjugate in the presence of 40 mM
NaCl.
Figure S44. Temperature dependent fluorescence spectra of Zn-aA conjugate in the presence of 40 mM
NaCl.
S22
Figure S45. Temperature dependent fluorescence spectra of Zn-aA conjugate in the presence of 100 mM
NaCl.
Figure S46. Temperature dependent fluorescence spectra of Zn-aA conjugate in the presence of 100 mM
NaCl.
S23
RLS studies of the DNA-porphyrin conjugates
Temperature dependent resonance light scattering (RLS) experiments were carried out in 1 mM Na-
cacodylate buffer, pH= 7.0 with different concentrations of NaCl. Temperature changing rate was 1 oC/min. The excitation and emission slits were both 2.5 nm.
Figure S47. Temperature dependent (cooling from 60 °C to -2 °C) RLS spectra of 2H-aA conjugate in
the presence of 0, 20, 40, and 100 mM NaCl, respectively.
S24
Figure S48. Temperature dependent (cooling from 60 °C to -2 °C) RLS spectra of Zn-aA conjugate in
the presence of 0, 40, and 100 mM NaCl, respectively.
S25
CD studies of the DNA-porphyrin conjugates
Figure S49. Temperature dependent (cooling from 50 °C to -2 °C) CD spectra of 2H-aA conjugate in the
presence of 0 and 20 mM NaCl, respectively.
Figure S50. Temperature dependent (cooling from 50 °C to -2 °C) CD spectra of Zn-aA conjugate in the
presence of 0, 20 and 40 mM NaCl, respectively.
S26
Figure S51. Temperature dependent (cooling from 50 °C to -2 °C) CD spectra of 2H-aT conjugate in the
presence of 0 and 100 mM NaCl, respectively.
Figure S52. Temperature dependent (cooling from 50 °C to -2 °C) CD spectra of 2H-pA conjugate in the
presence of 0 and 100 mM NaCl, respectively.
S27
Figure S53. Temperature dependent (cooling from 50 °C to -2 °C) CD spectra of 2H-pT conjugate in the
presence of 0 and 100 mM NaCl, respectively.
Figure S54. Temperature dependent (cooling from 50 °C to -2 °C) CD spectra of Zn-aT conjugate in the
presence of 0 and 100 mM NaCl, respectively.
S28
Figure S55. Temperature dependent (cooling from 50 °C to -2 °C) CD spectra of Zn-pA conjugate in the
presence of 0 and 100 mM NaCl, respectively.
Figure S56. Temperature dependent (cooling from 50 °C to -2 °C) CD spectra of Zn-pT conjugate in the
presence of 0 and 100 mM NaCl, respectively.
S29
Figure S57. The intensity of the induced CD signal in the porphyrin Soret band region at -2 °C as a
function of NaCl concentration (from 0 mM to 100 mM).
0 20 40 60 80 100
-2
0
2
4
6
8
10
12
CD
/ m
de
g[NaCl] / mM
2H-aA
Zn-aA
2H-pA
2H-pT
Zn-pA
Zn-pT
2H-aT
Zn-aT